Previously, it was established that H3-and C14-labeled chlorpromazine (CPZ) administered as a tracer to Rhesus monkeys chronically dosed with 20 mg/kg cold CPZ, was quantitatively excreted via the urine and feces over a period of three weeks. Chemical assay procedures for urinary drug derivatives demonstrated only about 60% of the urinary drug content, while 40% -- according to radioquantitation--remained in the aqueous phase after extraction of unconjugated CPZ metabolites and aglycones from conventional conjugates such as glucuronides and sulfates. The 40% residual radioactivity is derived from novel CPZ metabolities of unknown structure which do not yield quinoid radicals readily demonstrable by their intense color reactions with acids. Approximately one half of this fraction is hydrolyzable with Beta-glucosidase, yielding both known and unknown, but recognizable CPZ metabolities. Their structures will be studied during the upcoming third year of the study, during which essentially human urine will be investigated. The remaining 20% of the unknown urinary CPZ derivatives are extractable from the aqueous phase by tetrahydrofuran at pH 1, or may be converted into recognizable CPZ metabolities by strong acid hydrolysis (3 N HCl). All unknown CPZ metabolities will be investigated by high resolution NMR and GC/MS, and other biological media (blood, feces, saliva, CSF if available) will be analyzed especially with regard to occurrence of the novel metabolites. An automated GC/MS assay for CPZ and its metabolities extracted from biological media is being developed at this time, and analogous assays for piperazine-linked phenothiazines and structurally related tricyclic drugs are also scheduled during the third year of this project.